Hiding opaque eyes in transparent organisms: a potential role for larval eyeshine in stomatopod crustaceans.

Opaque screening pigments are a fundamental requisite for preserving resolution in image-forming eyes. Possession of any type of image-forming eye in a transparent, pelagic animal will thus undermine the ability of that animal to be invisible in the water column. Transparent, pelagic animals must therefore deal with the trade-off between the ability to see and the ability of other animals to see them. Stomatopod larvae, like many transparent crustaceans, possess specialized optics in their compound eyes that minimize the volume of the opaque retina. Though the volumes of these retinas are reduced, their opacity remains conspicuous to an observer. The light reflected from structures overlying the retinas of stomatopod crustacean larval eyes, referred to here as eyeshine, is hypothesized to further reduce the visibility of opaque retinas. Blue or green wavelengths of light are most strongly reflected in stomatopod larval eyeshine, suggesting a putative spectral matching to the light environment against which the larval eyes are viewed. We tested the efficacy of stomatopod crustacean larval eyeshine as an ocular camouflaging mechanism by photographing larvae in their natural light environment and analysing the contrast of eyes with the background light. To test for spectral matching between stomatopod larval eyeshine and the background light environment, we characterized the spectrum of eyeshine and calculated its performance using radiometric measurements collected at the time of each photographic series. These results are the first to demonstrate an operative mirror camouflage matched in both spectrum and radiance to the pelagic background light environment.

The molecular genetics and evolution of colour and polarization vision in stomatopod crustaceans.

Stomatopod crustaceans have the most complex assemblage of visual receptor classes known; retinas of many species are thought to express up to 16 different visual pigments. Physiological studies indicate that stomatopods contain up to six distinct middle-wavelength-sensitive (MWS) photoreceptor classes, suggesting that no more than six different MWS opsin gene copies exist per species. However, we previously reported the unexpected expression of 6-15 different MWS genes in retinas of each of five stomatopod species (Visual Neurosci 26: 255-266, 2009). Here, we present a review of the results reported in this publication, plus new results that shed light on the origins of the diverse colour and polarization visual capabilities of stomatopod crustaceans. Using in situ hybridization of opsins in photoreceptor cells, we obtained new results that support the hypothesis of an ancient functional division separating spatial and polarizational vision from colour vision in the stomatopods. Since evolutionary trace analysis indicates that stomatopod MWS opsins have diverged both with respect to spectral tuning and to cytoplasmic interactions, we have now further analyzed these data in an attempt to uncover the origins, diversity and potential specializations among clades for specific visual functions. The presence of many clusters of highly similar transcripts suggests exuberant opsin gene duplication has occurred in the stomatopods, together with more conservative, ancient gene duplication events within the stem crustacean lineage. Phylogenetic analysis of opsin relatedness suggests that opsins specialized for colour vision have diverged from those devoted to polarization vision, and possibly motion and spatial vision.

Tidal vertical migration: an endogenous rhythm in estuarine crab larvae.

Field-caught larvae of the estuarine crab Rhithropanopeus harrisii have a tidal rhythm of vertical migration when maintained in constant conditions. Laboratory-reared larvae do not show this rhythm. Endogenous tidal vertical migrations aid the retention of these planktonic larvae in estuaries near the parent populations.

Behavioural evidence for polarisation vision in stomatopods reveals a potential channel for communication.

Polarisation sensitivity (PS) - the ability to detect the orientation of polarised light - occurs in a wide variety of invertebrates [1] [2] and vertebrates [3] [4] [5], many of which are marine species [1]. Of these, the crustacea are particularly well documented in terms of their structural [6] and neural [7] [8] adaptations for PS. The few behavioural studies conducted on crustaceans demonstrate orientation to, or local navigation with, polarised sky patterns [9]. Aside from this, the function of PS in crustaceans, and indeed in most animals, remains obscure. Where PS can be shown to allow perception of polarised light as a 'special sensory quality' [1], separate from intensity or colour, it has been termed polarisation vision (PV). Here, within the remarkable visual system of the stomatopod crustaceans (mantis shrimps) [10], we provide the first demonstration of PV in the crustacea and the first convincing evidence for learning the orientation of polarised light in any animal. Using new polarimetric [11] and photographic methods to examine stomatopods, we found striking patterns of polarisation on their antennae and telson, suggesting that one function of PV in stomatopods may be communication [12]. PV may also be used for tasks such as navigation [5] [9] [13], location of reflective water surfaces [14] and contrast enhancement [1] [15] [16] [17] [18]. It is possible that the stomatopod PV system also contributes to some of these functions.

Photosensitivity spectrum of crayfish rhodopsin measured using fluorescence of metarhodopsin.

Discrepancies exist among spectral measurements of sensitivity of crayfish photoreceptors, their absorption in situ, and the number and absorption spectra of crayfish photopigments that are extracted by digitonin solutions. We have determined the photosensitivity spectrum of crayfish rhodopsin in isolated rhabdoms using long wavelength fluorescence emission from crayfish metarhodopsin as an intrinsic probe. There is no measurable metarhodopsin in the dark-adapted receptor, so changes in the emission level are directly proportional to metarhodopsin concentration. We therefore used changes in metarhodopsin fluorescence to construct relaxation and saturation ("photoequilibrium") spectra, from which the photosensitivity spectrum of crayfish rhodopsin was calculated. This spectrum peaks at or approximately 530 nm and closely resembles the previously measured difference spectrum for total bleaches of dark-adapted rhabdoms. Measurements of the kinetics of changes in rhabdom fluorescence and in transmittance at 580 nm were compared with predictions derived from several model systems containing one or two photopigments. The comparison shows that only a single rhodopsin and its metarhodopsin are present in the main rhabdom of crayfish, and that other explanations must be sought for the multiple pigments seen in digitonin solution. The same analysis shows that there is no detectable formation of isorhodopsin in the rhabdom.

Dark regeneration of rhodopsin in crayfish photoreceptors.

The eyes of crayfish were exposed to lights of known spectral composition, and the course of regeneration was followed in the dark by measuring the content of rhodopsin and metarhodopsin in single rhabdoms isolated at various times after the adaptation, using an assay that is based on the fluorescence of metarhodopsin. Complete recovery requires several days in the dark after intense adaptation to orange light, but requires less than 2 d after blue light exposure. Following an orange light exposure with blue produces recovery kinetics characteristic of the blue light exposure alone. This quickening of recovery occurs whether the receptors are exposed to blue light either immediately or many hours after the original exposure to orange. Conversely, following blue light adaptation with orange leads to slow recovery, which is characteristic of orange alone. Recovery from long-wavelength adaptation is slower principally because many rhabdoms seem to delay the onset of regeneration. We suggest that the regeneration system is itself photosensitive, and after orange light adaptation the supply of active chromophore (presumably 11-cis retinal) limits the rate of recovery. Once started, recovery proceeds slowly and continuously, and the total pigment concentration (rhodopsin plus metarhodopsin) in the rhabdomeric membrane remains approximately constant. Within hours after intense adapting exposures, the rhabdoms become altered in appearance, the surfaces become coated with accessory pigment, and the bands of microvilli are less distinct. These changes persist until recovery of rhodopsin proceeds, which suggests that visual pigment regeneration results from addition of newly synthesized rhodopsin associated with membrane turn-over.

Application of an invariant spectral form to the visual pigments of crustaceans: implications regarding the binding of the chromophore.

Visual pigment absorption spectra were measured in single photoreceptors of a stomatopod, a crayfish, a hermit crab, and five species of brachyuran crab. All fitted a Mansfield (1985) invariant form for visual pigment, the form also fitted by vertebrate retinal-based visual pigments. This is consistent with a theoretical model based on the structure of visual pigment molecules (Greenberg et al., 1975; Honig et al., 1976) which predicts that spectral bandwidth decreases as lambda max increases. The conformation to the invariant form implies that for any given chromophore bandwidth times lambda max is a constant.

Stomatopod photoreceptor spectral tuning as an adaptation for colour constancy in water.

Where colour is used in communication absolute judgement of signalling spectra is important, and failures of colour constancy may limit performance. Stomatopod crustaceans have unusual eyes in which the midband contains ten or more classes of photoreceptor. For constancy based on receptor adaptation to a fixed background, elementary theory predicts and we confirm by modelling, that stomatopods' narrow-band receptors outperform more broadly tuned receptors. Similar considerations could account for the small spectral separation of receptors in each midband row. Thus, stomatopods seem to trade-off sensitivity and signal-to-noise ratio for increased colour constancy.

Spectral tuning of dichromats to natural scenes.

Multispectral images of natural scenes were collected from both forests and coral reefs. We varied the wavelength position of receptors in hypothetical dichromatic visual systems and, for each receptor pair estimated the percentage of discriminable points in natural scenes. The optimal spectral tuning predicted by this model results in photoreceptor pairs very like those of forest dwelling, dichromatic mammals and of coral reef fishes. Variations of the natural illuminants in forests have little or no effect on optimal spectral tuning, but variations of depth in coral reefs have moderate effects on the spectral placement of S and L cones. The ratio of S and L cones typically found in dichromatic mammals reduces the discriminability of forest scenes; in contrast, the typical ratio of S and L cones in coral reef fishes achieves nearly the optimal discrimination in coral reef scenes.

Specialization of retinal function in the compound eyes of mantis shrimps.

Visual function and its specialization at the level of the retina were studied in 13 species of stomatopod crustaceans, representing three superfamilies: Gonodactyloidea, Lysiosquilloidea, and Squilloidea. We measured attenuation and irradiance spectra in the environment of each species, at the actual depths and times of activity where we observed individuals. We also characterized the intrahabdomal filters of all study species and determined the absolute spectral sensitivity functions and approximate photon capture rates of all photoreceptor classes below the level of the 8th retinular cell in seven of these species. Shallow-water gonodactyloid species have four distinct classes of intrarhabdomal filters, producing photoreceptors that are relatively insensitive but which have the broadest spectral coverage of all. Deep-water gonodactyloids and all lysiosquilloids have filters that are spectrally less diverse. These species often discard the proximal filter classes of one or more receptor types. As a result, their retinas are more sensitive but have reduced spectral range or diversity. The single squilloid species has the most sensitive photoreceptors of any we observed, due to the lack both of intrarhabdomal filters and tiered photoreceptors. Photon absorption rates, at the times of animal activity, were similar in most photoreceptor classes of all species, whether the receptors were tiered or untiered, or filtered or unfiltered. Thus, the retinas of stomatopods are specialized to operate at similar levels of stimulation at the times and depths of actual use, while evidently maintaining the greatest possible potential for spectral coverage and discrimination.

The intrarhabdomal filters in the retinas of mantis shrimps.

The intrarhabdomal filters in the photoreceptors of compounds eyes of 32 species of mantis shrimps (Crustacea: Stomatopoda), representing seven families within the superfamilies Lysiosquilloidea and Gonodactyloidea, were surveyed by microspectrophotometry of filters in fresh cryosections of the retina. A total of up to four classes of filters exist in stomatopods: two each in Rows 2 and 3 of the midband. All lysiosquilloid species lacked the proximal filter in Row 3; a few also lacked the proximal Row 2 class. While most gonodactyloid species had all four possible classes, in some species the proximal filters of Row 3 and (in one case) Row 2 were missing. In all, at least 11 distinct spectral classes of pigments were found. Absorption spectra suggested that the filter pigments were probably carotenoids or carotenoproteins, although the distal filter of Row 3 was often exceptional, appearing to contain a mixture of pigments. While the types of pigments found in the filters of the various species generally followed taxonomic lines, numerous exceptions were found that were apparently related to the ecological requirements of the various species.

Corneal curvatures and refractions of central American frogs.

We employed neutralizing infrared videophotorefraction and photokeratometry to examine the manifest refractions and corneal curvatures of 21 species of anurans (frogs and toads) in five families (Dendrobatidae, Bufonidae, Centrolenidae, Leptodactylidae, and Hylidae) resident in Central America. We found that all of the anurans exhibited hyperopic refractions in air, but that the observed hyperopia was not totally explained by the small eye artefact (Glickstein & Millodot, 1970). An allometric comparison of the corneal radii of these small anurans with those of a large number of other vertebrates, inferred from ocular axial lengths, showed that their corneal radii increased significantly more rapidly with increasing body size than that of other vertebrates generally (allometric slope constants: anurans: 0.270 +/- 0.032; other vertebrates: 0.151 +/- 0.004). Among the anurans examined, nocturnal Hylids had significantly larger eyes than diurnal Dendrobatid frogs and Bufonid toads.

Ultraviolet photoreception in mantis shrimp.

An UV-sensitive class of photoreceptors exists in all regions of the retinas of mantis shrimps. UV photosensitivity apparently resides in rhabdomeres of the eighth retinular cell (R8) that lies atop each rhabdom; and in ommatidia where the R8 rhabdomere consists of microvilli parallel in a single direction, sensitivity is maximal when the e-vector of plane-polarized light is parallel to the microvilli. Spectral sensitivity of the UV photoreceptor peaks at 345 nm and is best explained by the presence of a photopigment with lambda max near 325 nm overlain by material that absorbs UV light at wavelengths below approximately 350 nm. Rhabdomeres of R8 cells in several different retinal regions of a variety of species examined contain a photopigment absorbing maximally below 340 nm. Under appropriate conditions, a metapigment with lambda max near 460 nm can be formed. UV vision may be useful for enhancing the visual contrast of midwater predators or prey.

Parallel processing and image analysis in the eyes of mantis shrimps.

The compound eyes of mantis shrimps, a group of tropical marine crustaceans, incorporate principles of serial and parallel processing of visual information that may be applicable to artificial imaging systems. Their eyes include numerous specializations for analysis of the spectral and polarizational properties of light, and include more photoreceptor classes for analysis of ultraviolet light, color, and polarization than occur in any other known visual system. This is possible because receptors in different regions of the eye are anatomically diverse and incorporate unusual structural features, such as spectral filters, not seen in other compound eyes. Unlike eyes of most other animals, eyes of mantis shrimps must move to acquire some types of visual information and to integrate color and polarization with spatial vision. Information leaving the retina appears to be processed into numerous parallel data streams leading into the central nervous system, greatly reducing the analytical requirements at higher levels. Many of these unusual features of mantis shrimp vision may inspire new sensor designs for machine vision.

Shrimps that pay attention: saccadic eye movements in stomatopod crustaceans.

Discovering that a shrimp can flick its eyes over to a fish and follow up by tracking it or flicking back to observe something else implies a 'primate-like' awareness of the immediate environment that we do not normally associate with crustaceans. For several reasons, stomatopods (mantis shrimp) do not fit the general mould of their subphylum, and here we add saccadic, acquisitional eye movements to their repertoire of unusual visual capabilities. Optically, their apposition compound eyes contain an area of heightened acuity, in some ways similar to the fovea of vertebrate eyes. Using rapid eye movements of up to several hundred degrees per second, objects of interest are placed under the scrutiny of this area. While other arthropod species, including insects and spiders, are known to possess and use acute zones in similar saccadic gaze relocations, stomatopods are the only crustacean known with such abilities. Differences among species exist, generally reflecting both the eye size and lifestyle of the animal, with the larger-eyed more sedentary species producing slower saccades than the smaller-eyed, more active species. Possessing the ability to rapidly look at and assess objects is ecologically important for mantis shrimps, as their lifestyle is, by any standards, fast, furious and deadly.

Visual Rhythms in Stomatopod Crustaceans Observed in the Pseudopupil.

Many aspects of visual function in animals are influenced by the operation of endogenous rhythms. Using techniques of intracellular optical physiology, I investigated visual rhythms in two species of stomatopod crustaceans (mantis shrimps): Squilla empusa, a species active throughout the day and night, and Gonodactylus oerstedii, which is strictly diurnal. Reflectance from within the deep pseudopupil of the compound eyes and its change upon stimulation with light were monitored in individual animals in constant conditions for up to two weeks. Both species expressed circadian rhythms in visual function. In S. empusa, the pupillary response was much stronger during subjective night; little or no response could be elicited during subjective day. In this species, an endogenous rhythm caused pupillary reflectance to increase during subjective day. Rhythms in G. oerstedii were of lower amplitude than in S. empusa and were more difficult to detect. The differences between these species, together with the results of other comparative research on visual rhythms in arthropods, suggest that circadian, rhythmic processes are involved in optimizing nocturnal eyes for maximum sensitivity and dynamic range.

The retinoids of seven species of mantis shrimp.

Eyes of stomatopod crustaceans, or mantis shrimps, contain the greatest diversity of visual pigments yet described in any species, with as many as ten or more spectral classes present in a single retina. In this study, the eyes of seven species of mantis shrimp from three superfamilies of stomatopods were examined for their content of retinoids. Only retinal and retinol were found; neither hydroxyretinoids nor dehydroretinoids were detected. The principal isomers were 11-cis and all-trans. The eyes of most of these species contain stores of 11-cis retinol, principally as retinyl esters, and in amounts in excess of retinal. Squilla empusa is particularly noteworthy, with over 5000 pmoles of retinol per eye.

The molecular evolution of visual pigments of freshwater crayfishes (Decapoda: Cambaridae).

This study examines the diverse maximum wavelength absorption (lambdamax) found in crayfishes (Decapoda: Cambaridae and Parastacidae) and the associated genetic variation in their opsin locus. We measured the wavelength absorption in the photoreceptors of six species that inhabit environments of different light intensities (i.e., burrows, streams, standing waters, and subterranean waters). Our results indicate that there is relatively little variation in lambdamax (522-530 nm) among species from different genera and families. The existing variation did not correlate with the habitat differences of the crayfishes studied. We simultaneously sequenced the rhodopsin gene to identify the amino acid replacements that affect shifts in maximum wavelength absorption. We then related these to changes that correlated with shifts in lambdamax by reconstructing ancestral character states using a maximum-likelihood approach. Using amino acid sequences obtained from five species (all were 301 amino acids in length), we identified a number of candidates for producing shifts of 4 to 8 nm in lambdamax. These amino acid replacements occurred in similar regions to those involved in spectral shifts in vertebrates.

Polarization contrast vision in Octopus.

While the ability to analyze polarized light is widespread among animals, its contribution to form vision has not yet been documented. We tested the hypothesis that polarization vision can be used for object discrimination, by training octopuses to distinguish between targets on the basis of the presence or absence of a pattern produced by a 90 degrees polarization contrast within the target. Octopuses recognized a 90 degrees contrast pattern within a single target, when presented either on a horizontal/vertical axis or on a 45 degrees/135 degrees axis. They were able to transfer their learning to new situations and to detect a polarization contrast when the orientations of the e-vector of light passing through the target center and background differed by as little as 20 degrees. Polarization vision may provide information similar to that available from color vision and thus serve to enhance the detection and recognition of objects.